Visual primer for identifying carcasses of silky fowl and black-bone silky fowl, kit and detection method therefor
A technology for silkie and black-bone chickens, applied in biochemical equipment and methods, measurement/testing of microbes, DNA/RNA fragments, etc., can solve the problem that there is no effective way to identify silky silkie and black-bone chicken carcasses and other issues, to achieve good social value, broad market application prospects, and simple operation
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Embodiment 1
[0033] Example 1 Verification of Genome Differences between Silky Chicken and Black Phoenix Chicken
[0034] Step 1: Template Preparation
[0035] (1) Tissue collection: The skin samples of 24 silky chickens and 24 black-bone chickens were collected.
[0036] (2) DNA template preparation: about 50 mg of skin samples were collected, put into a centrifuge tube with 500 μL of liquid nitrogen, and the collected samples were ground with a tissue grinder. After the liquid nitrogen was evaporated to dryness, 200 microliters of TE was added, centrifuged at 1200 rpm / min, and the supernatant was extracted for later use.
[0037] Step 2: PCR Amplification
[0038] (1) PCR amplification reaction system
[0039] PCR amplification primers are shown in Table 1:
[0040] Table 1 PCR amplification primers
[0041]
[0042] The PCR reaction system is shown in Table 2:
[0043] Table 2 PCR amplification system
[0044] Reagent Volume (ul) 2x reaction buffer 12.5 ...
Embodiment 2
[0048] Reliability of Embodiment 2 Single-blind Detection and Judgment Method
[0049] In this example, a single-blind method is adopted to determine the source of the species of the samples of known species according to the molecular detection results.
[0050] Step 1: Template Preparation
[0051] (1) Tissue collection: The skin samples of 12 silky chickens and 12 black-bone chickens were collected.
[0052] (2) DNA template preparation: about 50 mg of skin samples were collected, put into a centrifuge tube with 500 μL of liquid nitrogen, and the collected samples were ground with a tissue grinder. After the liquid nitrogen was evaporated to dryness, 200 microliters of TE was added, centrifuged at 1200 rpm / min, and the supernatant was extracted for later use.
[0053] Step 2: PCR Amplification
[0054] (1) PCR amplification reaction system
[0055] PCR amplification primers are shown in Table 3:
[0056] Table 3 PCR amplification primers
[0057]
[0058] The PCR re...
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